Ultraviolet (UV) radiation was one of the primary contributors to skin aging. Prolonged exposure to UV rays accelerated the degradation of collagen, resulting in various photoaging-related skin problems, including wrinkles, dryness, and hyperpigmentation. Mesotherapy was a widely utilized skin repair method in the field of aesthetic medicine; however, it faced challenges such as biosafety concerns and monofunctionality. Herein, we developed for the first time a highly bioactive yak collagen mesotherapy (Col-M) for the rejuvenation of photoaging skin. Through experiments measuring DPPH and ABTS radical scavenging rates, tyrosinase inhibition rates, and zebrafish efficacy evaluation experiments, the multifunctional Col-M was identified. This mesotherapy demonstrated exceptional antioxidant, whitening, moisturizing, repair-promoting, and soothing effects. Animal studies demonstrated that the Col-M increased dermal density, improved skin tone, enhanced skin hydration, and restored transcutaneous water loss (TEWL). The Col-M stimulated fibroblasts, providing an optimal physiological environment that promoted collagen regeneration and improved photoaged skin. H&E staining revealed that Col-M effectively avoided triggering an inflammatory response while demonstrating the ability to suppress epidermal hyperplasia. Masson trichrome staining illustrated that the Col-M promoted collagen regeneration and expedited the remodeling process of photoaged skin. In photoaged mice skin, Col-M elevated hydroxyproline content, superoxide dismutase (SOD) activity, and glutathione reductase (GSH) levels, concurrently reducing malondialdehyde (MDA) levels. The Col-M provided an effective treatment for photoaged skin, presenting significant application potential in the fields of dermatology and regenerative medicine.